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Plasmid Files

pAc-sgRNA-Cas9

Plasmid for expressing a single guide RNA (sgRNA) together with Cas9 in Drosophila cells.

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pAc-sgRNA-Cas9 Sequence and MappAc-sgRNA-Cas9.dna
Map and Sequence File   
Sequence Author:  Liu Lab / Addgene #49330
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 polIII terminator SapI (85) SapI (106) ATG SV40 NLS pAc-sgRNA-Cas9 10,663 bp
SapI  (85)
2 sites
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
SapI  (106)
2 sites
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
Cas9
3109 .. 7209  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
3109 .. 7209  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Ac5 promoter
507 .. 2948  =  2442 bp
Drosophila melanogaster actin 5c promoter
Ac5 promoter
507 .. 2948  =  2442 bp
Drosophila melanogaster actin 5c promoter
AmpR
9473 .. 10,333  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   9473 .. 10,264  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
9473 .. 10,333  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   10,265 .. 10,333  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
9473 .. 10,333  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
7351 .. 7950  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
7351 .. 7950  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
8714 .. 9302  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
8714 .. 9302  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
dU6 promoter
107 .. 506  =  400 bp
RNA polymerase III promoter for Drosophila U6-2
snRNA (Das et al., 1987)
dU6 promoter
107 .. 506  =  400 bp
RNA polymerase III promoter for Drosophila U6-2
snRNA (Das et al., 1987)
SV40 poly(A) signal
7958 .. 8092  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7958 .. 8092  =  135 bp
SV40 polyadenylation signal
AmpR promoter
10,334 .. 10,438  =  105 bp
AmpR promoter
10,334 .. 10,438  =  105 bp
gRNA scaffold
13 .. 88  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
13 .. 88  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
3xFLAG
2992 .. 3057  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
3xFLAG
2992 .. 3057  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
T2A
7273 .. 7326  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid
protein
Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides.
T2A
7273 .. 7326  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid
protein
Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides.
nucleoplasmin NLS
7210 .. 7257  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
7210 .. 7257  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
SV40 NLS
3064 .. 3084  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
3064 .. 3084  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
polIII terminator
7 .. 12  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
7 .. 12  =  6 bp
RNA polymerase III transcription terminator
ATG
2989 .. 2991  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2989 .. 2991  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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